Overview

  • Product name
    Anti-Estrogen Receptor alpha antibody [TE111.5D11]
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Mouse monoclonal [TE111.5D11] to Estrogen Receptor alpha
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, Neutralisingmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment corresponding to Human Estrogen Receptor alpha aa 302-595. Recombinant fragment, corresponding to amino acids 302-595 of Human Estrogen Receptor alpha.
    Database link: P03372

  • Positive control
    • MCF-7 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab16460 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

EMSA 1/1.
IP Use at an assay dependent concentration. Use at a concentration of 2 µg/mg of protein lysate on native and denatured lysate.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 66 kDa).
Neutralising Use at an assay dependent concentration. This antibody can be used to block binding of estrogen to the estrogen receptor.

Target

  • Function
    Nuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Can activate the transcriptional activity of TFF1.
  • Sequence similarities
    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Domain
    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • Post-translational
    modifications
    Phosphorylated by cyclin A/CDK2. Phosphorylation probably enhances transcriptional activity.
    Glycosylated; contains N-acetylglucosamine, probably O-linked.
    Ubiquitinated. Deubiquitinated by OTUB1.
    Dimethylated by PRMT1 at Arg-260. The methylation may favor cytoplasmic localization.
    Palmitoylated (isoform 3). Not biotinylated (isoform 3).
  • Cellular localization
    Nucleus. Cytoplasm. Cell membrane. A minor fraction is associated with the inner membrane and Nucleus. Cytoplasm. Cell membrane. Associated with the inner membrane via palmitoylation.
  • Information by UniProt
  • Database links
  • Alternative names
    • DKFZp686N23123 antibody
    • ER alpha antibody
    • ER antibody
    • ER-alpha antibody
    • Era antibody
    • ESR antibody
    • ESR1 antibody
    • ESR1_HUMAN antibody
    • ESRA antibody
    • Estradiol receptor antibody
    • Estrogen nuclear receptor alpha antibody
    • Estrogen receptor 1 antibody
    • Estrogen receptor alpha 3*,4,5,6,7*/822 isoform antibody
    • Estrogen receptor alpha antibody
    • Estrogen receptor alpha delta 3*,4,5,6,7*,8*/941 isoform antibody
    • Estrogen receptor alpha delta 3*,4,5,6,7*/819 2 isoform antibody
    • Estrogen receptor alpha delta 4 +49 isoform antibody
    • Estrogen receptor alpha delta 4*,5,6,7*/654 isoform antibody
    • Estrogen receptor alpha delta 4*,5,6,7,8*/901 isoform antibody
    • Estrogen receptor alpha E1 E2 1 2 antibody
    • Estrogen receptor alpha E1 N2 E2 1 2 antibody
    • Estrogen receptor antibody
    • ESTRR antibody
    • NR3A1 antibody
    • Nuclear receptor subfamily 3 group A member 1 antibody
    see all

Images

  • ab16460 at 1µg/ml detecting Estrogen Recptor from MCF-7 cells by Western blot. ab16460 at 1µg/ml detecting Estrogen Recptor from MCF-7 cells by Western blot.
  • Overlay histogram showing MCF7 cells stained with ab16460 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16460, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References

This product has been referenced in:
  • Deng X  et al. Guanxin Danshen Formulation Protects against Myocardial Ischemia Reperfusion Injury-Induced Left Ventricular Remodeling by Upregulating Estrogen Receptor ß. Front Pharmacol 8:777 (2017). Read more (PubMed: 29163163) »
  • Elbaradie KB  et al. Sex-specific response of rat costochondral cartilage growth plate chondrocytes to 17ß-estradiol involves differential regulation of plasma membrane associated estrogen receptors. Biochim Biophys Acta 1833:1165-72 (2013). Read more (PubMed: 23305904) »
See all 4 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Answer

Gracias por tu respuesta.

No hay ninguna recomendación específica acerca de la concentración usada como bloqueante para este anticuerpo. La concentración usada depende totalmente de las condiciones experimentales, y debe ser calculada por el usuario.

Siento no poder ser de más ayuda. Si tienes cualquier otra consulta, por favor, no dudes en contactarme de nuevo.

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Question
Answer

Thank you for your call.

I can confirm that immunogen for this antibody was a recombinant fragment corresponding to amino acids 302-595 of Human Estrogen Receptor alpha.

This sequence show a 55% homology to Human Estrogen Receptor beta and we therefore would not expect a cross reaction of the antibody. But this was not experimentally tested and therefore is not guaranteed.

I hope this information is helpful and wish you a good weekend.

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Question
Answer

Thank you for your reply.
I am not sure if these amino acids correspond with beta estradiol, however the immunogen that was used to generate the antibody is a recombinant fragment, corresponding to amino acids 302-595 of Human Estrogen Receptor. Therefore you could align this section of the human estrogen receptor with your target sequence, and see the level of similarity.
Please let me know if there is anything else I can help you with.

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Answer

Thank you for contacting Abcam.
The antibody ab16460 will bind to the estorgen receptor and the immunogen is a recombinant fragment, corresponding to amino acids 302-595 of Human Estrogen Receptor.
If there is anything else I can help you with, please let me know.

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Answer

Thank you for sending the customer's western blot image. I am having difficulty reading it, and seeing where the signal is detected in the blots. I understand that the customer expects to see the receptor at 65 kDa, but does not. Can you please ask the customer: 1. What are the samples (species, cell types)? How are they prepared (RIPA lysis buffer?) and are they collected with protease inhibitors? Does the customer boil the samples to denature, with a reducing agent? 2. How are the membranes blocked? With 5% milk? 3. What is the dilution or concentration of the antibody that the customer applies tot he membrane? 4. What are the sizes (molecular weights) of the bands that the customer does detect? I look forward to resolving this issue for your customer.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (MCF7, Monocytes and Monocyte-Derived-Macrophages)
Loading amount
25 µg
Specification
MCF7, Monocytes and Monocyte-Derived-Macrophages
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Apr 16 2008

Answer

Thank you for your enquiry. I would like to recommend the following ER alpha antibody; Estrogen Receptor alpha antibody [33] (ab2746). we have good feedback regarding the behaviour of this antibody by western blotting. It is also a monoclonal antibody. We do not have any knowledge of non-specific reactivity towards ER beta. I have also performed an alignment of the immunogen used with the ER beta sequence and there is only very minimal sequence similarity suggesting that this antibody would not show this kind of reactivity. With regards beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) we do not have specific details of MCF7 detection by this antibody. However, I would certainly expect it to detect beta actin in this cell type using an HRP conjugated secondary antibody. Furthermore we would guarantee it for this purpose. Please see our "Abpromise". I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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